scholarly journals Development of an in-house Lepto-LAT kit for screening and detection of Leptospira infection

2021 ◽  
Vol 19 (3) ◽  
pp. 529-537
Author(s):  
Le Thi Lan Anh ◽  
Vu Thi Thuong ◽  
Pham Thi Ha Giang ◽  
Bui Thi Thanh Nga ◽  
Minh Thi Hang ◽  
...  

Leptospirosis is a common zoonotic disease in the tropics and subtropics. Leptospira infected human without prompt detection and treatment will face serious consequences such as acute hepatitis-kidneys, pulmonary hemorrhage which can lead to death. Besides the MAT gold standard method, Leptosipra antigens developed by DNA recombination technology have been widely studying and applying in diagnosis of Leptospira infection in human and animal. Overcoming the disadvantages of MAT and ELISA such as complicated protocol, which requires highly qualified staff and specialized equipment, the latex agglutination method has been studied and widely used in detecting pathogens such as Salmonella, E. coli, Leptospira in the world. The advantages of this method are simple operation, fast and cheap. In the previous article, we expressed Leptospira LigB antigen in E. coli cells and successfully purified it by affinity chromatography with 98% purity. In this paper, we present the process for establishment of a Lepto-LAT kit to detect Leptospira infection in dogs. This kit had the sensitivity and specificity of 91.75% and 91.57%, respectively.

2005 ◽  
Vol 68 (10) ◽  
pp. 2208-2211 ◽  
Author(s):  
NIRMALA THAMPURAN ◽  
A. SURENDRARAJ ◽  
P. K. SURENDRAN

Escherichia coli is a common contaminant of seafood in the tropics and is often encountered in high numbers. The count of E. coli as well as verotoxigenic E. coli O157:H7 was estimated in 414 finfish samples composed of 23 species of fresh fish from retail markets and frozen fish from cold storage outlets in and around Cochin, India. A total of 484 presumptive E. coli were isolated, and their indole–methyl red–Voges-Proskauer–citrate (IMViC) pattern was determined. These strains were also tested for labile toxin production by a reverse passive latex agglutination method and checked for E. coli serotype O157 by latex agglutination with O157-specific antisera. Certain biochemical marker tests, such as methylumbelliferyl-β-glucuronide (MUG), sorbitol fermentation, decarboxylase reactions, and hemolysis, which are useful for screening pathogenic E. coli, were also carried out. Results showed that 81.4% of the E. coli isolates were sorbitol positive. Among this group, 82% were MUG positive, and 14.46% of the total E. coli isolates showed human blood hemolysis. None of the isolates were positive for agglutination with E. coli O157 antisera nor did any produce heat-labile enterotoxin. This study indicates that typical E. coli O157 or labile toxin–producing E. coli is absent in the fish and fishery environments of Cochin (India). However, the presence of MUG and sorbitol-negative strains that are also hemolytic indicates the existence of aberrant strains, which require further investigation.


2020 ◽  
Vol 14 (4) ◽  
pp. 312-324
Author(s):  
Sadra S. Tehrani ◽  
Abolfazl Jahangiri ◽  
Mortaza Taheri-Anganeh ◽  
Hossein Maghsoudi ◽  
Saeed Khalili ◽  
...  

Background: Cholera triggered by Vibrio cholerae remains the main reason for morbidity and mortality all over the world. In addition, salmonellosis is regarded as an infectious disease that makes it essential for the identification and detection of Salmonella. With a beta-barrel structure consisting of eight non-parallel beta strands, OmpW family is widely distributed among gram-negative bacteria. Moreover, OmpW isolated from S. typhimurium and Vibrio cholerae can be used in vaccine design. Methods: Topology prediction was determined. T-cell and B-cell epitopes were selected from exposed areas, and sequence conservancy was evaluated. The remaining loops and inaccessible residues were removed to prepare OmpW-1. High antigenicity peptides were detected to replace inappropriate residues to obtain OmpW-2. Physicochemical properties were assessed, and antigenicity, hydrophobicity, flexibility, and accessibility were compared to the native Omp-W structure. Low score areas were removed from the designed structure for preparing the OmpW-3. To construct OmpW-4, TTFrC was used as T-CD4+ cell-stimulating factor and CTB as adjuvant to the end of the C-terminal of this sequence, which can increase the antigenicity and sequence density. The sequences were re-analyzed to delete the unfavorable residues. Besides, the solubility of the mature OmpW and the designed structure were predicted while overexpressed in E. coli. Results: The designed vaccine is a stable protein which has immune cells recognizing epitopes and is considered as an antigen. The construct can be overexpressed in a E. coli. Conclusion: The multi-epitope vaccine is a suitable stimulator for immune system and would be a candidate for experimental research. Recent patents describing numerous inventions related to the clinical facets of vaccine peptide against human infectious disease.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Lan N. Truong ◽  
Brayden D. Whitlock

AbstractControlling infections has become one of the biggest problems in the world, whether measured in lives lost or money spent. This is worsening as pathogens continue becoming resistant to therapeutics. Antimicrobial surfaces are one strategy being investigated in an attempt to decrease the spread of infections through the most common route of transmission: surfaces, including hands. Regulators have chosen two hours as the time point at which efficacy should be measured. The objectives of this study were to characterize the new antimicrobial surface compressed sodium chloride (CSC) so that its action may be understood at timepoints more relevant to real-time infection control, under two minutes; to develop a sensitive method to test efficacy at short time points; and to investigate antifungal properties for the first time. E. coli and Candida auris are added to surfaces, and the surfaces are monitored by contact plate, or by washing into collection vats. An improved method of testing antimicrobial efficacy is reported. Antimicrobial CSC achieves at least 99.9% reduction of E. coli in the first two minutes of contact, and at least 99% reduction of C. auris in one minute.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Femi Ayoade ◽  
Judith Oguzie ◽  
Philomena Eromon ◽  
Omolola E. Omotosho ◽  
Tosin Ogunbiyi ◽  
...  

AbstractShiga toxigenic strains of E. coli (STEC) known to be etiological agents for diarrhea were screened for their incidence/occurrence in selected abattoirs sources in Osogbo metropolis of Osun State, Nigeria using a randomized block design. Samples were plated directly on selective and differential media and E. coli isolates. Multiplex PCR analysis was used to screen for the presence of specific virulence factors. These were confirmed serologically as non-O157 STEC using latex agglutination serotyping kit. Sequence analysis of PCR products was performed on a representative isolate showing the highest combination of virulence genes using the 16S gene for identification purposes only. Results showed that the average cfu/cm2 was significantly lower in the samples collected at Sekona-2 slaughter slab compared with those collected at Al-maleek batch abattoir and Sekona-1 slaughter slab in ascending order at P = 0.03. Moreover, the average cfu/cm2E. coli in samples collected from butchering knife was significantly lower when compared with that of the workers’ hand (P = 0.047) and slaughtering floor (P = 0.047) but not with the slaughter table (P = 0.98) and effluent water from the abattoir house (P = 0.39). These data suggest that the abattoir type may not be as important in the prevalence and spread of STEC as the hygiene practices of the workers. Sequence analysis of a representative isolate showed 100% coverage and 96.46% percentage identity with Escherichia coli O113:H21 (GenBank Accession number: CP031892.1) strain from Canada. This sequence was subsequently submitted to GenBank with accession number MW463885. From evolutionary analyses, the strain from Nigeria, sequenced in this study, is evolutionarily distant when compared with the publicly available sequences from Nigeria. Although no case of E. coli O157 was found within the study area, percent occurrence of non-O157 STEC as high as 46.3% at some of the sampled sites is worrisome and requires regulatory interventions in ensuring hygienic practices at the abattoirs within the study area.


2021 ◽  
Vol 14 (2) ◽  
pp. 78-90
Author(s):  
Ahmed Jarad ◽  
Kh. Al- Jeboori

The present study focus on non-O157 Shiga toxin-producing E. Coli (STEC), included a bacteriological study was subjected to provide additional information for non-O157 STEC prevalence in children and calves. Isolation by using selective culturing media (CHROMagar STEC and CHROMagar O157) from 127 children suffering from diarrhea and 133 calves in Al- Muthanna province. Characterization depends on culturing positive colony on MacConkey agar and Levin’s Eosin Methylene blue agar, staining single colony from the growth by gram stain, biochemical tests; Indole, the Methyl Red, Voges-Proskauer, Citrate test, Oxidase, Catalase, Urease, Motility, Kligler Iron and Api-20E, were done to confirm a diagnosis of non-O157 STEC, The reliable isolation as non-O157 STEC serotyping by specific latex agglutination test for the target non-O157 STEC (big six) serogroup (O26, O45, O103, O111, O121 and O145). The current study showed the prevalence of non-O157 STEC was 20 of out 127 (15.73%) in samples collected from children and 27 / 133 (20.30%) in calves samples in conclusion the Non-O157 STEC is an important cause of diarrhea in children, and calves; finally, the calves play an important reservoir for Non-O157 STEC.


Author(s):  
Maryna Nochka ◽  

The article is devoted to the analysis tools for assessing human capital based on world rankings in the context of sustainable development. The most famous world rankings of human capital, studied by such international organizations as the World Bank, the United Nations, the World Economic Forum, the University of Groningen in collaboration with the University of California at Davis and others, are considered. Quantifying human capital as the economic and social value of a skill set is measured through an index. Each organization makes measurements according to its own method. The application of different criteria and indicators for assessing human capital at the macroeconomic level is analyzed. The considered assessment methodologies are overwhelmingly based on statistical approaches. Analyzed the position of Ukraine in the world rankings in recent years in dynamics. It has been confirmed that these international ratings can be considered as a reflection of the state of human capital in Ukraine. Revealed quite high rating positions of Ukraine in comparison with other countries. The results allow us to conclude that there is insufficient government funding for the development of human capital. It is concluded that Ukraine needs to improve the quality of human capital as a leading factor in increasing the efficiency of the country's economy in the context of sustainable development. The study showed that the use of high-quality, highly qualified human capital leads to an improvement in the country's position in the world rankings.


1994 ◽  
Vol 57 (4) ◽  
pp. 337-340 ◽  
Author(s):  
AJIBOLA O. FAPOHUNDA ◽  
KENNETH W. MCMILLIN ◽  
DOUGLAS L. MARSHALL ◽  
W. M. WAITES

Isolates of Escherichia coli and Clostridium perfringens from beef and Aeromonas hydrophila from fish were examined for their ability to survive and grow as cross-contaminates on nonnative tissues at simulated ambient (35°C) and aging/conditioning (15°C) temperatures of handling and retailing found in the tropics. Growth of all isolates over a 10-h period was greater (P < 0.05) on their native tissues at both temperatures. The aging/conditioning temperature effectively limited growth of E. coli and A. hydrophila to less than l-logl0 CFU/g and prevented growth of C. perfringens on beef and fish samples. All three isolates demonstrated characteristic mesophilic growth response on both tissues at 35°C during the 10-h retail period. The study suggests that two muscle food products could be jointly handled to efficiently use available storage/haulage capacity in tropical countries. Potential savings in space, labor and energy would be made if cross-contamination between the two products is minimized by available packaging and sanitizing technologies.


2018 ◽  
Vol 40 (4) ◽  
pp. 366-374 ◽  
Author(s):  
Fellype de Carvalho Barreto ◽  
Cleber Rafael Vieira da Costa ◽  
Luciene Machado dos Reis ◽  
Melani Ribeiro Custódio

Abstract Renal osteodystrophy (ROD), a group of metabolic bone diseases secondary to chronic kidney disease (CKD), still represents a great challenge to nephrologists. Its management is tailored by the type of bone lesion - of high or low turnover - that cannot be accurately predicted by serum biomarkers of bone remodeling available in daily clinical practice, mainly parathyroid hormone (PTH) and alkaline phosphatase (AP). In view of this limitation, bone biopsy followed by bone quantitative histomorphometry, the gold-standard method for the diagnosis of ROD, is still considered of paramount importance. Bone biopsy has also been recommended for evaluation of osteoporosis in the CKD setting to help physicians choose the best anti-osteoporotic drug. Importantly, bone biopsy is the sole diagnostic method capable of providing dynamic information on bone metabolism. Trabecular and cortical bones may be analyzed separately by evaluating their structural and dynamic parameters, thickness and porosity, respectively. Deposition of metals, such as aluminum and iron, on bone may also be detected. Despite of these unique characteristics, the interest on bone biopsy has declined over the last years and there are currently few centers around the world specialized on bone histomorphometry. In this review, we will discuss the bone biopsy technique, its indications, and the main information it can provide. The interest on bone biopsy should be renewed and nephrologists should be capacitated to perform it as part of their training during medical residency.


2020 ◽  
Vol 27 (07) ◽  
pp. 1363-1370
Author(s):  
Aneela Khawaja ◽  
Iffat Javed ◽  
Sohaila Mushtaq ◽  
Saeed Anwar ◽  
Faiqa Arshad ◽  
...  

Antimicrobial resistance (AMR) is a devastating question that is threatening the health globally. The extensive and indiscriminative use of antibiotics has evolved a notorious resistance in Staphylococcus aureus.  This resistance developed through possession of mecA gene, which codes for modified penicillin binding protein (PBP2a) and the emergent strain being labeled “methicillin resistant Staphylococcus aureus”. Conventional phenotypic techniques for detection of MRSA rely on standardization of cultural characteristics. The drawbacks of diagnostic error to report MRSA include: poor prognosis, expensive treatment, dissemination of multi-drug resistant strains and even treatment failure. Latex agglutination method can be adopted as a more accurate and quick strategy for rapid detection of methicillin resistance. Objectives: To compare detection of mecA gene in methicillin resistant isolates of Staphylococcus aureus by latex agglutination and PCR; by assessing the sensitivity and specificity of both methods. Study Design: Descriptive Cross-Sectional study. Setting: Pathology Department, Post Graduate Medical Institute, Lahore. Period: From January 2015 to December 2015; according to standard operating procedures at Microbiology laboratory. Material & Methods: A total 713 consecutive, non-duplicate isolates of Staphylococcus aureus were processed. Methicillin resistance was determined using cefoxitin (30mg) by Kirby-Bauer method using CLSI guideline (2016), latex agglutination method; and PCR for mecA gene. Results: The results showed that out of 713 Staphylococcus aureus isolates, 92 (12.90%) isolates were resistant to cefoxitin and were labelled as MRSA. majority MRSA isolates recovered from pus (44.57%) and wound swab (20.65%), followed by blood (13.04%), fluid (8.70%), CSF (4.35%), CVP (3.26%), HVS (3.26%) and tracheal secretion (2.17%). By latex agglutination method, 87 (94.50%) were positive for PBP2a; while on PCR mecA gene was detected only in 82 (89.10%) MRSA isolates. When assessed with PCR (gold standard) the sensitivity and diagnostic accuracy of latex agglutination was 100% and 94.57%, respectively. Conclusion: Latex agglutination test can be employed as rapid and reliable diagnostic technique in MRSA isolates for mecA gene detection, where resources for molecular methods are inadequate. This can effectually lessen the misdiagnosis of resistant strains, and over/ ill-use of antibiotics.


Author(s):  
R. O. Chioma ◽  
I. Mzungu ◽  
J. B. Orpin

Escherichia coli infections and poor nutritional status have implications on the growth and development of children under five years, physically, mentally and health wise with consequences such as diarrhoea, stunting, wasting, underweight and often times leading to death, depending on their severity. This study evaluated the antibiogram of Escherichia coli O157 and Verocytotoxigenic Escherichia coli (VTEC)and the nutritional status of diarrhoeic children under five years in Kaduna State, Nigeria, using Conventional isolation methods, latex agglutination tests, VTEC-ELISA tests, Chi-square (SPSS Version 19) and WHO Antro (Version 3.2.2). Purposive sampling was used to select 350 children presenting with diarrhoea in six government hospitals within the three senatorial zones of Kaduna State. The results obtained revealed that 76(21.7%) of the 350 stool samples were positive for E. coli and 28(36.8%) were positive for E. coli O157:H7serotype and 1(1.3%) verocytotoxigenic E. coli (VTEC) serotype. High susceptibility to ciprofloxacin, chloramphenicol and high resistance to sulphamethoxazole, cefotaxime, amoxicillin, gentamicin and tetracycline by the isolates were observed. The study concluded that antibiotics have not been very effective in the treatment of E. coli-related diarrhoea, with VTEC now emerging in this part of the world, making it a serious public health issue. The study therefore recommends the implementation of programmes geared towards good hygiene, good nutrition and good health.


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